IL-10 predicts incident neuroinflammatory disease and proviral load dynamics in a large Brazilian cohort of people living with human T-lymphotropic virus type 1.

Human T-Lymphotropic Virus type-1 (HTLV-1) is a unique retrovirus associated with both leukemogenesis and a specific neuroinflammatory condition known as HTLV-1-Associated Myelopathy (HAM). Currently, most proposed HAM biomarkers require invasive CSF sampling, which is not suitable for large cohorts or repeated prospective screening. To identify non-invasive biomarkers for incident HAM in a large Brazilian cohort of PLwHTLV-1 (n=615 with 6,673 person-years of clinical follow-up), we selected all plasma samples available at the time of entry in the cohort (between 1997-2019), in which up to 43 cytokines/chemokines and immune mediators were measured. Thus, we selected 110 People Living with HTLV-1 (PLwHTLV-1), of which 68 were neurologically asymptomatic (AS) at baseline and 42 HAM patients. Nine incident HAM cases were identified among 68 AS during follow-up. Using multivariate logistic regression, we found that lower IL-10, IL-4 and female sex were independent predictors of clinical progression to definite HAM (AUROC 0.91), and outperformed previously suggested biomarkers age, sex and proviral load (AUROC 0.77). Moreover, baseline IL-10 significantly predicted proviral load dynamics at follow-up in all PLwHTLV-1. In an exploratory analysis, we identified additional plasma biomarkers which were able to discriminate iHAM from either AS (IL6Rα, IL-27) or HAM (IL-29/IFN-λ1, Osteopontin, and TNFR2). In conclusion, female sex and low anti-inflammatory IL-10 and IL-4 are independent risk factors for incident HAM in PLwHTLV-1,while proviral load is not, in agreement with IL-10 being upstream of proviral load dynamics. Additional candidate biomarkers IL-29/IL-6R/TNFR2 represent plausible therapeutic targets for future clinical trials in HAM patients.

OPRM1 and BDNF polymorphisms associated with a compensatory neurophysiologic signature in knee osteoarthritis patients.

OBJECTIVE: The present study investigated the relationship between three genetic polymorphisms of OPRM1 (rs1799971 - A118G and rs1799972 - C17T) and BDNF (rs6265 - C196T) and EEG-measured brain oscillations in Knee Osteoarthritis (KOA) patients. MATERIALS AND METHODS: We performed a cross-sectional analysis of a cohort study (DEFINE cohort), KOA arm, with 66 patients, considering demographic (age, sex, and education), clinical (pain intensity and duration), OPRM1 (rs1799971 - A118G and rs1799972 - C17T) and BDNF (rs6265 - C196T) genotypes, and electrophysiological measures. Brain oscillations relative power from Delta, Theta, Alpha, Low Alpha, High Alpha, Beta, Low Beta and High Beta oscillations were measured during resting state EEG. Multivariate regression models were used to explore the main brain oscillation predictors of the three genetic polymorphisms. RESULTS: Our findings demonstrate that Theta and Low Beta oscillations are associated with the variant allele of OPRM1-rs1799971 (A118G) on left frontal and left central regions, respectively, while Alpha brain oscillation is associated with variant genotypes (CT/TT) of BDNF-rs6265 on frontal (decrease of oscillation power) and left central (increase of oscillation power) regions. No significant model was found for OPRM1-rs1799972 (C17T) in addition to the inclusion of pain intensity as a significant predictor of this last model. CONCLUSION: One potential interpretation for these findings is that polymorphisms of OPRM1 - that is involved with endogenous pain control - lead to increased compensatory oscillatory mechanisms, characterized by increased theta oscillations. Along the same line, polymorphisms of the BDNF lead to decreased alpha oscillations in the frontal area, likely also reflecting the disruption of resting states to also compensate for the increased injury associated with knee OA. It is possible that these polymorphisms require additional brain adaption to the knee OA related injury.

Association of Mu opioid receptor (A118G) and BDNF (G196A) polymorphisms with rehabilitation-induced cortical inhibition and analgesic response in chronic osteoarthritis pain.

Background/objective: Chronic pain due to osteoarthritis (OA) is a prevalent cause of global disability. New biomarkers are needed to improve treatment allocation, and genetic polymorphisms are promising candidates. Method: We aimed to assess the association of OPRM1 (A118G and C17T) and brain-derived neurotrophic factor (BDNF [G196A]) polymorphisms with pain-related outcomes and motor cortex excitability metrics (measured by transcranial magnetic stimulation) in 113 knee OA patients with chronic pain. We performed adjusted multivariate regression analyses to compare carriers versus non-carriers in terms of clinical and neurophysiological characteristics at baseline, and treatment response (pain reduction and increased cortical inhibitory tonus) after rehabilitation. Results: Compared to non-carriers, participants with polymorphisms on both OPRM1 (A118G) and BDNF (G196A) genes were less likely to improve pain after rehabilitation (85 and 72% fewer odds of improvement, respectively). Likewise, both carriers of OPRM1 polymorphisms (A118G and C17T) were also less likely to improve cortical inhibition (short intracortical inhibition [SICI], and intracortical facilitation [ICF], respectively). While pain and cortical inhibition improvement did not correlate in the total sample, the presence of OPRM1 (A118G) and BDNF (G196A) polymorphisms moderated this relationship. Conclusions: These results underscore the promising role of combining genetic and neurophysiological markers to endotype the treatment response in this population.

The copy number variation of GSTM1 as a promising prognostic factor of oral squamous cell carcinoma.

OBJECTIVE: The aim of this study was to investigate whether the copy number variation (CNV) of GSTM1 and GSTT1 is related to the occurrence of oral squamous cell carcinoma (OSCC) relapses, along the overall and progression-free survival of patients. STUDY DESIGN: A total of 234 OSCC patients were recruited from the Heliópolis hospital and they were distributed among 4 groups according to the occurrence of OSCC relapses. Fisher exact test, odds ratio (OR), and 95% CI were determined to investigate the chances of OSCC progression. The overall and progression-free survival were analyzed by the Kaplan-Meier and Cox regression methods. RESULTS: The CNV of GSTM1 analysis showed that one copy of the gene was associated with reduced chances of OSCC recurrences (OR 0.45; 95% CI 0.25-0.81) and decreased the risk of tumor progression (HR 0.50; 95% CI 0.33-0.75). Furthermore, one copy of GSTM1 was related to a better overall survival rate (HR 0.63; 95% CI 0.0.44-0.91). Regarding the CNV of GSTT1, no copies were associated with the chances of OSCC relapses, the overall survival, or the progression-free survival. CONCLUSIONS: The CNV of GSTM1 may be applied to predict OSCC relapses and aid the treatment management, which might improve the survival rates of patients.

Casework direct kit as an alternative extraction method to enhance touch DNA samples analysis.

Latent fingerprints are commonly found in crime scenes, and currently used in forensic analysis to obtain STR profiles from DNA recovered from finger contact. Analysis of STR profiles obtained from touch DNA has been very useful to elucidate crimes and the extraction method may be determinant for the recovery of genetic material collected from different surfaces. This study aimed to verify and compare the efficiency of two different extraction kits for processing touch DNA samples obtained from fingerprints deposited on computer keyboards, knife handles and exterior door handles and steering wheels of cars. One hundred and four experiments were conducted to simulate crime scenes and evaluate the efficiency of two extraction kits for touch DNA samples: the DNA IQ™ System and the Casework Direct Kit (both Promega Corporation). Each experiment was conducted with two individuals in order to obtain a mixture profile. The genetic material deposited was collected by double swab method (Sweet et al. 1997) and DNA quantification was conducted using Quantifiler Trio™ (ThermoFisher Scientific). Samples were amplified by PowerPlex® Fusion System kit (Promega). It was possible to obtain STR profiles for 32 (61.5%) out of the 52 extracted using DNA IQ and 51 (98.1%) out of the 52 extracted using the Casework Direct Kit. Samples extracted by DNA IQ had higher average of quantification values for long targets (>200bp) across all tested surfaces. That seems to be due to an incompatibility between the Quantifiler Trio and the Casework Direct Kit. Samples with positive quantification but without STR profile, as well as samples without quantification but with STR profiles were also observed. Statistical analysis showed that the Casework Direct Kit produced significantly more useful profiles than DNA IQ (p-value = 0.001), since these profiles had more STR markers with allelic correspondence to second donators present in the mixture. This study provides insights about the effect of different surfaces and extraction methods on recovery and generation of STR profiles. Limitations for the quantification step for these samples with a low quantity of DNA were highlighted as well. We concluded that the Casework Direct Kit was much more efficient for processing touch DNA samples than DNA IQ.

Polymorphisms of CHRNA3 and CHRNA5: Head and neck cancer and cigarette consumption intensity in a Brazilian population.

BACKGROUND: Cigarette consumption has been identified as the main non-etiological factor in head and neck cancer (HNC) development. One of the main compounds in cigarettes is nicotine, which binds directly to nicotine acetylcholine receptors (nAchRs) in the body, which are encoded by different genes of the CHRNA family. Polymorphisms in some of these genes have been studied in relation to the risk of HNC and cigarette consumption intensity. The aim of this study was to evaluate whether there were associations between the CHRNA3 (rs578776) and CHRNA5 (rs16969968) polymorphisms and HNC risk and between the polymorphisms and the intensity of cigarette consumption. METHODS: A total of 1,067 individuals from Heliopolis Hospital in São Paulo were investigated, including 619 patients with HNC and 448 patients without diagnosed tumors. All participants answered a questionnaire about sociodemographic information and cigarette consumption data. The polymorphisms were determined by TaqMan genotyping by real-time PCR. RESULTS: The polymorphisms studied, rs578776 (CHRNA3) and rs16969968 (CHRNA5), did not have an association with HNC risk, but the rs16969968 homozygous genotype was associated with increased cigarette consumption intensity (OR 1.93, 95% CI 1.05-3.58). CONCLUSION: The polymorphism CHRNA5 can be considered an indirect risk factor for neoplasms in these Brazilian samples when cigarette consumption increased.

Alterations of the CCND1 and HER-2/neu (ERBB2) proteins in esophageal and gastric cancers.

We evaluated the relationship of amplification and polysomy of both the CCND1 and the ERBB2 (alias HER-2/NEU) genes to the overexpression of their proteins in esophageal and gastric cancers and also their association with clinicopathological features. CCND1 gene amplification (45%) was more prevalent than polysomy (25%) in esophageal carcinoma, but the pattern observed was similar in gastric adenocarcinoma (10% amplification, 15% polysomy). For ERBB2, polysomy was a more frequent mechanism than amplification in both esophageal (32.5 vs. 7.5%) and gastric (15 vs. 5%) cancers. Overexpression of cyclin D1 protein was identified in 37.5% of the specimens of esophageal tumors and 35% of gastric tumors, and overexpression of Her-2/neu protein in 12.5 and 7.5%, respectively. The kappa-statistics revealed a fair agreement in both types of tumors only in overexpression and amplification of the CCND1 gene; the ERBB2 gene showed a fair agreement in amplification and polysomy and the level of protein expression in gastric adenocarcinoma. Thus, polysomy 17 could contribute to a high Her-2/neu protein level, at least in gastric cancer. Our data indicated an association with alcohol consumption and the CCND1 gene or protein levels, in both esophageal and gastric cancers.

IL28B gene polymorphism SNP rs8099917 genotype GG is associated with HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) in HTLV-1 carriers.

BACKGROUND: The polymorphisms of IL28B have been described as important in the pathogenesis of infections caused by some viruses. The aim of this research was to evaluate whether IL28B gene polymorphisms (SNP rs8099917 and SNP rs12979860) are associated with HAM/TSP. METHODS: The study included 229 subjects, classified according to their neurological status in two groups: Group I (136 asymptomatic HTLV-1 carriers) and Group II (93 HAM/TSP patients). The proviral loads were quantified, and the rs8099917 and rs12979860 SNPs in the region of IL28B-gene were analyzed by StepOnePlus Real-time PCR System. RESULTS: A multivariate model analysis, including gender, age, and HTLV-1 DNA proviral load, showed that IL28B polymorphisms were independently associated with HAM/TSP outcome in rs12979860 genotype CT (OR = 2.03; IC95% = 0.96-4.27) and in rs8099917 genotype GG (OR = 7.61; IC95%  = 1.82-31.72). CONCLUSION: Subjects with SNP rs8099917 genotype GG and rs12979618 genotype CT may present a distinct immune response against HTLV-1 infection. So, it seems reasonable to suggest that a search for IL28B polymorphisms should be performed for all HTLV-1-infected subjects in order to monitor their risk for disease development; however, since this is the first description of such finding in the literature, we should first replicate this study with more HTLV-1-infected persons to strengthen the evidence already provided by our results.

Establishment and characterization of androgen-independent human prostate cancer cell lines, PcBra1, PcBra2, and PcBra3.

One of the main obstacles for understanding biological events involved in cancer is the lack of experimental models for in vitro studies especially for prostate cancer (PC). There are a limited number of PC cell lines being the majority originated from metastatic tumors mostly acquired from American Tissue Cell Culture which demands importation an expensive and bureaucratic process. Also it is well known that there are ethnic differences between populations concerning the behavior of tumors and the research based on cell lines derived from Brazilians should be interesting. Our aim was to develop tumor cell lines from primary PC.

Genes de reparo do DNA e de susceptibilidade genética em pacientes com melanoma maligno / DNA repair and genetic susceptibility genes in malignant melanoma patients

O melanoma é uma lesão maligna da pele, com alta taxa de mortalidade cuja incidência vem aumentando nos últimos anos. Os principais fatores de risco são a história familial da doença, presença de nevos benignos múltiplos ou nevos atípicos e melanoma prévio. Imunossupressão, sensibilidade ao sol e exposição intermitente e intensa à radiação UV da luz solar, sem proteção, são fatores de risco adicionais. O objetivo deste estudo caso-controle de base hospitalar foi avaliar a contribuição de polimorfismos de genes de metabolização de xenobióticos (CYP1A1/MspI, CYP2E1/PstI, GSTM1, GSTT1 e GSTP1/Bsma), de genes de reparo do DNA (XRCC1/MspI, XRCC3/NcoI e XPD/PstI) e do gene do receptor de vitamina D (VDR/FokI e VDR/TaqI) no risco de melanoma. Consentiram em participar 193 pacientes com melanoma (49,7% homens e 50,3% mulheres, média de 52 ± 14,28 anos) e 208 controles (51,4% homens e 48,6% mulheres, média de 48 ± 15,24 anos) que após responderem a um questionário detalhado sobre hábitos e tipos de exposição a fatores de risco cederam amostras biológicas para análsie do DNA por PCR-RFLP. Os principais fatores de risco para o melanoma foram ascendência européia (p<0,001), cor de olhos claros (p<0,001), presença de nevos (p<0,001), histórico de queimadura grave na adolescência (p<0,001), falta de filtro solar (p<0.034) e exposição à lâmpadas fluorescentes (p=0,001). Quanto à análise dos polimorfismos de genes de metabolização de xenobióticos somente o GSTT1 nulo revelou associação inversamente positiva com o risco de melanoma maligno (OR ajustado = 0,60; IC95% = 0,37-0,97). Entretanto, essa associação não se manteve após a análise de regressão múltipla escalonada. Os polimorfismos VDR/FokI e VDR/TaqI não modificaram a susceptibilidade ao melanoma maligno na comparação entre os grupos...

Melanoma is a malignant skin lesion, with high mortalitty rate and its incidence has been rising in the last years. The main risk factors are melanoma family history, presence of multiple benign or atypical nevi and previous melanoma. Immunosuppression, sun sensitivy and intermittent and intense exposure to UV sunlight radiation, without protection, are additional risk factors. The aim of this hospital based case-control study was to evaluate the contribution of genetic polymorphisms of xenobiotic metabolizing enzymes (CYP1A1/MspI, CYP2E1/PstI, GSTM1, GSTT1 and GSTP1/Bsma), DNA repair genes (XRCC1/MspI, XRCC3/NcoI and XPD/PstI) and vitamin D receptor genes (VDR/FokI and VDR/TaqI) to the risk of melanoma. All participants, including 193 melanoma patients (49.7% men and 50.3% women, mean age 52 ± 14.28 years old) and 208 controls (51.4% men and 48.6% women, mean age 48 ± 15.24 years old) gave written informed consent to participate in the study and agreed to donate a sample of bloody to analysis of DNA by PCR-RFLP and answer a questionarie regarding phenotypic characteristics, personal habits and questions regarding sun exposure that could be associated to the disease. The main risk factors to melanoma were European ancestries (p<0.001), light colored eyes (p<0.001), presence of nevi (p<0.001), history of sunburns during the adolescence (p<0.001), no use of sunblock (p<0.034) and exposure of fluorescent lamps (p<0.001). Regarding the genes polymorphisms, only GSTT1 null genotype showed as an inversely positivefactor (OR adjusted = 0.60; 95%CI = 0.37-0.97) to malignant melanoma. However, this association disappeared with multiple regression analysis. The VDR/FokI and VDR/TaqI polymorphisms did not alter the susceptibility to malignant melanoma in the comparison between groups...